A binding site for Pax proteins regulates expression of the gene for the neural cell adhesion molecule in the embryonic spinal cord
A binding site for Pax proteins regulates expression of the gene for the neural cell adhesion molecule in the embryonic spinal cord
复制标题
DOI：
10.1073/pnas.94.4.1465
复制
发表时间：
1997-02-18
影响因子：
11.1
通讯作者：
Edelman, GM
中科院分区：
文献类型：
--
作者：
Holst, BD;Wang, YB;Edelman, GM
神经细胞粘附分子（N-CAM）介导细胞间相互作用，并在发育过程中以特征性时空模式表达。在之前对控制N-CAM基因表达的因素的研究中，我们确定了Pax蛋白配对结构域的结合位点（指定为PBS）中。在该研究中，我们证明了已知对中枢神经系统发育重要的转录因子Pax-6与N-CAM PBS结合，并表明PBS可影响体内N-CAM表达。在COS-1细胞中产生的Pax-6通过两个半位点PBS-1和PBS-2与PBS结合;这两个位点的突变完全破坏了结合。此外，来自胚胎日龄（E）11.5的小鼠胚胎的核提取物与PBS结合，并且这种结合被Pax-6的抗体抑制。为了确定PBS在体内的作用，我们产生了具有N-CAM启动子/lacZ基因构建体的转基因小鼠，所述构建体含有野生型或突变的PBS。PBS-1和PBS-2中的突变降低了脊髓套层中β-半乳糖苷酶表达的程度，将其限制在E11.5、E14.5、这些突变消除了在野生型脊髓中观察到的大部分表达。结合我们先前的PBS结合多种Pax蛋白的观察，数据表明这种结合有助于神经发育期间N-CAM基因表达的调节。
The neural cell adhesion molecule (N-CAM) mediates cell-cell interactions and is expressed in characteristic spatiotemporal patterns during development, In previous studies of factors that control N-CAM gene expression, we identified a binding site for the paired domain of Pax proteins (designated PBS) in the mouse N-CAM promoter, In this study, we demonstrate that a transcription factor known to be important for development of the central nervous system, Pax-6, binds to the N-CAM PBS and show that the PBS can influence N-CAM expression in vivo. Pax-6, produced in COS-1 cells, bound to the PBS through two half-sites, PBS-1 and PBS-2; mutations in both of these sites completely disrupted binding, Moreover, nuclear extracts from embryonic day (E) 11.5 mouse embryos bound to the PBS, and this binding was inhibited by antibodies to Pax-6. To determine the role of the PBS in vivo, we generated transgenic mice with N-CAM promoter/lacZ gene constructs containing either a wild-type or a mutated PBS, Mutations in PBS-1 and PBS-2 decreased the extent of beta-galactosidase expression in the mantle layer of the spinal cord limiting it to ventral regions at E11.5, At E14.5, these mutations eliminated most of the expression that was seen in the wild-type spinal cord, Taken together with our previous observations that the PBS binds multiple Pax proteins, the data indicate that such binding contributes to the regulation of N-CAM gene expression during neural development.